ES2280217T3 - IL-8 RECEIVER ANTAGONISTS. - Google Patents

Abstract

A compound selected from the group consisting of: 4 - [[3- (2-bromophenyl) -4-oxo-1- (phenylmethyl) -2-imidazolidinylidene] imino] -3-hydroxybenzonitrile; 4 - [[(7aS) -2- (2-bromophenyl) -hexahydro-1-oxo-3H-pyrrolo [1,2-c] imidazol-3-ylidene] amino] -3-hydroxybenzonitrile; 4 - [[5- (2-bromophenyl) -3, 3a, 4, 5-tetrahydro-1, 1-dioxide-4-oxoimidazo [1,5-b] isothiazol-6 (2H) -iliden] amino] - 3-hydroxybenzonitrile; 4 - [[(6R, 7aS) -2- (2-bromophenyl) -hexahydro-6-hydroxy-1-oxo-3H-pyrrolo [1,2-c] imidazol-3-ylidene] amino] -3-hydroxybenzonitrile ; 4 - [[(7aR) -2- (2-bromophenyl) -hexahydro-1-oxo-3 H -pyrrolo [1,2-c] imidazol-3-ylidene] amino] -3-hydroxybenzonitrile; 4 - [(S) -2- (2, 3-Dichloro-phenyl) -1-oxo-hexahydro-pyrrolo [1,2-c] imidazol-3-ylidenamino] -3-hydroxy-benzonitrile; 4 - [(R) -2- (2, 3-Dichloro-phenyl) -1-oxo-hexahydro-pyrrolo [1,2-c] imidazol-3-ylidenamino] -3-hydroxy-benzonitrile; 4- [2- (2, 3-Dichloro-phenyl) -1-oxo-hexahydro-pyrrolo [1,2-c] imidazol-3-ylidenamino] -3-hydroxy-benzonitrile; 4 - [(S) -2- (2-Bromo-phenyl) -1-oxo-hexahydro-pyrrolo [1,2-c] imidazol-3-ylidenamino] -3-hydroxy-benzonitrile; 4 - [(R) -2- (2-Bromo-phenyl) -1-oxo-hexahydro-pyrrolo [1,2-c] imidazol-3-ylidenamino] -3-hydroxy-benzonitrile; 4- [2- (2-Bromo-phenyl) -1-oxo-hexahydro-pyrrolo [1,2-c] imidazol-3-ylidenamino] -3-hydroxy-benzonitrile, and; 4 - [[2- (2-bromophenyl) -1, 5, 6, 7, 8, 8a-hexahydro-1-oxoimidazo [1,5-a] pyridin-3 (2H) -iliden] amino] -3- hydroxybenzonitrile; or a pharmaceutically acceptable salt thereof.

Description

Receptor antagonists IL-8

Field of the Invention

This invention relates to a new group of guanidine compounds, to procedures for the preparation of same, to their use in the treatment of diseases mediated by IL-8, GRO?, GRO?, GRO γ, NAP-2, and ENA-78 and a pharmaceutical compositions for use in said therapy.

Background of the invention

Many different names have been applied to Interleukin-8 (IL-8), such as attraction-active protein-1.
tion of neutrophils (NAP-1), monocyte-derived neutrophil chemotactic factor (MDNCF), neutrophil activating factor (NAF), and chemoattractant factor of T-cell lymphocytes. Interleukin-8 is a chemoattractant for neutrophils, basophils, and a subset of T cells. It is produced by a large number of nucleated cells including macrophages, fibroblasts, endothelial and epithelial cells exposed to TNF, IL-1?, IL-1? or LPS, and neutrophils themselves when exposed to LPS or chemotactic factors such as FMLP. M. Baggiolini et al ., J. Clin. Invest . 84, 1045 (1989); J. Schroder et al ., J. Immunol . 139, 3474 (1987) and J. Immunol . 144, 2223 (1990); Strieter, et al ., Science 243, 1467 (1989) and J. Biol. Chem . 264, 10621 (1989); Cassatella et al ., J. Immunol . 148, 3216 (1992).

GRO?, GRO?, GRO? And NAP-2 also belong to the family of chemokines?. Like IL-8, these chemokines have also been called by different names. For example, GRO?,?,? Have been designated MGSA?,? And? Respectively (Melanoma Growth Stimulating Activity), see Richmond et al ., J. Cell Physiology 129, 375 (1986) and Chang et al ., J. Immunol 148, 451 (1992). All chemokines of the α family that possess the ELR motif directly preceding the CXC motif bind to the IL-8 B receptor.

IL-8, GRO?, GRO?, GRO?, NAP-2 and ENA-78 stimulate numerous functions in vitro . All of them have been shown to have chemoattractant properties for neutrophils, while IL-8 and GROα have demonstrated chemotactic activity against T lymphocytes, and basophils. In addition, IL-8 can induce the release of histamine from basophils of normal and atopic individuals. GRO-? And IL-8 can also induce the release of lysozomal enzymes and the respiratory outbreak of neutrophils. IL-8 has also been shown to increase surface expression of Mac-1 (CD11b / CD18) on neutrophils without de novo protein synthesis. This may contribute to increased adhesion of neutrophils to vascular endothelial cells. Many known diseases are characterized by a massive neutrophil infiltration. As IL-8, GRO?, GRO?, GRO? And NAP-2 promote the accumulation and activation of neutrophils, these chemokines have been implicated in a wide variety of acute and chronic inflammatory disorders including psoriasis and rheumatoid arthritis, Baggiolini et al ., FEBS Lett . 307, 97 (1992); Miller et al ., Crit. Rev. Immunol . 12, 17 (1992); Oppenheim et al ., Annu. Rev. Immunol . 9, 617 (1991); Seitz et al ., J. Clin. Invest . 87, 463 (1991); Miller et al ., Am. Rev. Respir. Pis . 146, 427 (1992); Donnely et al ., Lancet 341, 643 (1993). In addition, ELR chemokines (those containing amino acids with the ELR motif just before the CXC motif) have also been implicated in angiostasis, Strieter et al ., Science 258, 1798 (1992).

In vitro , IL-8, GRO?, GRO?, GRO?, And NAP-2 induce form change, chemotaxis, granule release, and neutrophil respiratory burst, binding and activating receptors of the family of seven transmembrane domains coupled to G proteins, in particular binding to the IL-8 receptors, more particularly the B receptor, Thomas et al ., J. Biol. Chem . 266, 14839 (1991); and Holmes et al ., Science 253, 1278 (1991). The development of non-peptide small molecule antagonists to become members of this family of receptors is unprecedented. For a report, see R. Freidinger in: Progress in Drug Research , Vol. 40, p. 33-98, Birkhauser Verlag, Basel 1993. Therefore, the IL-8 receptor represents a promising target for the development of new anti-inflammatory agents.

Two high affinity human IL-8 receptors (77% homology) have been characterized: IL-8Rα, which binds only to IL-8 with high affinity, and IL-8Rβ, which has high affinity for IL-8 as well as GRO-?, GRO?, GRO? And NAP-2. See Holmes et al ., Supra ; Murphy et al ., Science 253, 1280 (1991); Lee et al ., J. Biol. Chem . 267, 16283 (1992); LaRosa et al ., J. Biol. Chem . 267, 25402 (1992); and Gayle et al ., J. Biol. Chem . 268, 7283 (1993).

There is still a need for treatment, in this field, for compounds that are capable of binding to the receptor IL-8? Or?. Therefore, the conditions associated with an increase in the production of IL-8 (which is responsible for the chemotaxis of neutrophils and subsets of T cells at the inflammatory site) could benefit from compounds that are binding inhibitors to the IL-8 receptor.

Detailed description of the invention

The compounds of the invention can be used together with the veterinary treatment of mammals, other than humans, in need of inhibition of IL-8 or other chemokines that bind to the receptors of IL-8 RA and RB. Diseases mediated by chemokines for treatment, therapeutic or prophylactic, in Animals include pathologies such as those indicated in this document in the Treatment Procedures section.

The compounds of the present invention are:

4 - [[3- (2-Bromophenyl) -4-oxo-1- (phenylmethyl) -2-imidazolidinyliden] imino] -3-hydroxybenzonitrile;

4 - [[(7aS) -2- (2-bromophenyl) -hexahydro-1-oxo-3 H -pyrrolo [1,2-c] imidazol-3-ylidene] amino] -3-hydroxybenzonitrile;

4 - [[5- (2-bromophenyl) -3,3a, 4,5-tetrahydro-1,1-dioxide-4-oxoimidazo [1,5-b] isothiazol-6 (2H) -yliden] amino] - 3-hi-
droxibenzonitrile;

4 - [[(6R, 7aS) -2- (2-bromophenyl) -hexahydro-6-hydroxy-1-oxo-3H-pyrrolo [1,2-c] imidazol-3-ylidene] amino] -3-hydroxy -
benzonitrile;

4 - [[(7aR) -2- (2-bromophenyl) -hexahydro-1-oxo-3H-pyrrolo [1,2-c] imidazol-3-ylidene] amino] -3-hydroxybenzonitrile,

4 - [(S) -2- (2,3-Dichloro-phenyl) -1-oxo-hexahydro-pyrrolo [1,2-c] imidazol-3-ylidenamino] -3-hydroxy-benzonitrile,

4 - [(R) -2- (2,3-Dichloro-phenyl) -1-oxo-hexahydro-pyrrolo [1,2-c] imidazol-3-ylidenamino] -3-hydroxy-benzonitrile,

4- [2- (2,3-Dichloro-phenyl) -1-oxo-hexahydro-pyrrolo [1,2-c] imidazol-3-ylidenamino] -3-hydroxy-benzonitrile,

4 - [(S) -2- (2-Bromo-phenyl) -1-oxo-hexahydro-pyrrolo [1,2-c] imidazol-3-ylidenamino] -3-hydroxy-benzonitrile,

4 - [(R) -2- (2-Bromo-phenyl) -1-oxo-hexahydro-pyrrolo [1,2-c] imidazol-3-ylidenamino] -3-hydroxy-benzonitrile,

4- [2- (2-Bromo-phenyl) -1-oxo-hexahydro-pyrrolo [1,2-c] imidazol-3-ylidenamino] -3-hydroxy-benzonitrile, Y

4 - [[2- (2-Bromophenyl) -1,5,6,7,8,8a-hexahydro-1-oxoimidazo [1,5-a] pyridin-3 (2H) -iliden] amino] -3- hydroxybenzoni-
trilo; or a pharmaceutically acceptable salt thereof,

Suitable pharmaceutically acceptable salts they are well known to those skilled in the art and include salts basic inorganic and organic acids, such as acid hydrochloric, hydrobromic acid, sulfuric acid, phosphoric acid, methanesulfonic acid, ethanesulfonic acid, acetic acid, acid malic, tartaric acid, citric acid, lactic acid, acid oxalic acid, succinic acid, fumaric acid, maleic acid, acid benzoic, salicylic acid, phenylacetic acid and mandelic acid. In addition, pharmaceutically acceptable salts may also be formed. of compounds of the invention with a pharmaceutically cation acceptable, for example, if a substituent group comprises a moiety carboxy Suitable pharmaceutically acceptable cations those skilled in the art are well known and include cations of alkali, alkaline earth and quaternary ammonium metals.

Preparation Procedures

The compounds of the invention can be obtained applying synthesis procedures, some of which are illustrated in the following Schemes.

Once the core of the guanidine, other compounds of these formulas can be prepared applying conventional techniques for group interconversion functional, well known in the art.

Scheme one

one

Aniline 6-scheme-1 desired can be prepared from benzoxazolinone 1-scheme-1 available in the market. 2-scheme-1 bromide can be prepared from benzoxazolinone 1-scheme-1 using conditions conventional bromination such as bromine and sodium acetate in acetic acid. Bromide 2-scheme-1 can become the cyanide 3-scheme-1 using conventional procedures such as copper (I) cyanide in DMF at reflux. Amide 3-scheme-1 can become the compound 4-scheme-1 protected with BOC using  conventional conditions such as BOC anhydride and triethylamine with a catalytic amount of dimethylaminopyridine in methylene chloride or other suitable organic solvent. The oxazolinone 4-scheme-1 can become aniline 6-scheme-1 desired by hydrolyzing first to phenol 5-scheme-1 using conventional conditions such as potassium carbonate in methanol followed by removal of the BOC protecting group using conditions conventional such as trifluoroacetic acid in chloride methylene or other suitable organic solvent giving the aniline 6-scheme-1.

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Scheme 2

2

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Thiourea 3-scheme-2 desired can be prepared as shown in scheme 2. Aniline 1-scheme-2 can be coupled with a commercially available isothiocyanate or with an isothiocyanate made from the condensation of an amine available in the market with thiophosgene or a thiophosgene equivalent giving thiourea 2-scheme-2. Phenol 2-scheme-2 can be protected as the TBS ether 3-scheme-2 using conventional conditions such as TBSCl and imidazole in THF or another suitable organic solvent. As an alternative the amine 1-scheme-2 can become the 5-scheme-2 isothiocyanate first protecting the phenol 1-scheme-2 with a protective group suitable such as ether TBS 4-scheme-2 using conditions conventional such as TBSCl and an amine base such as imidazole in THF or other suitable organic solvent. Thiourea 5-scheme-2 can be prepared condensing the amine 4-scheme 2 with thiophosgene in presence of a base such as potassium carbonate. Thiourea 3-scheme-2 desired can be prepared from isothiocyanate 5-scheme-2 condensing it with the desired amine in a suitable organic solvent such as ethanol or DMF

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Scheme 3

3

Lactam 3-scheme-3 desired can be prepared as shown in Scheme 3. Carbodiimide 2-scheme-3 can be prepared to from thiourea 1-scheme-3 using conventional conditions such as an excess amount of methanesulfonyl chloride and a suitable amine base such as triethylamine in an organic solvent preferably methylene at room temperature. Lactam 3-scheme-3 can be prepared to from carbodiimide 2-scheme-3 condensing in first place carbodiimide 2-scheme-3 with the desired α-amino ester in a solvent suitable organic such as THF followed by withdrawal from the TBS group using conventional conditions such as TBAF in THF at 0 ° C.

Synthesis Examples

The invention will now be described by making reference to the following examples that are simply illustrative and should not be considered as a limitation of Scope of the present invention. All temperatures are given in degrees Celsius, all solvents are of the highest purity available and all reactions are carried out in conditions anhydrous in an argon atmosphere unless another is indicated thing.

In the Examples, all temperatures are at degrees Celsius (ºC). Mass spectra are performed in a VG Zab mass spectrometer using rapid atom bombardment, to unless otherwise indicated. The spectra of 1 H-NMR (hereinafter referred to as "NMR") were recorded at 250 MHz using a Bruker spectrometer AM 250 or Am 400. The multiplicities indicated are; s = singlet, d = doublet, t = triplet, c = quadruplet, m = multiplet and a indicates a wide signal Sat. indicates a saturated solution, eq. indicates the ratio of a molar equivalent of reagent to reagent principal.

Example 1 Preparation of 4 - [[3- (2-Bromophenyl) -4-oxo-1- (phenylmethyl) -2-imidazolidinyliden] imino] -3-hydroxybenzonitrile a) Preparation of 4-bromo-1,2-benzoxazolinone

To a solution of benzoxazolinone (10 g, 74 mmol) in acetic acid (50 ml) at 0 ° C sodium acetate was added (7.4 g, 74 mmol) and bromine (3.8 ml, 74 mmol) and the reaction mixture will warm to room temperature. After 21 h, the precipitate was collected by filtration and washed with water. The filtrate was concentrated under reduced pressure to give more material. solid that was collected by filtration followed by washing with water. He material was combined giving 14 g (88%) of 4-bromo-1,2-benzoxazolinone in the form of a yellow solid that did not need purification additional. 1 H NMR (400 MHz, DMSO) δ 7.6 (s, 1 H), 7.3 (d, 1 H), 7.0 (d, 1 H); MS (EI) m / e 212 (M +).

b) Preparation of 4-cyano-1,2-benzoxazolinone

To a solution of 4-bromo-1,2-benzoxazolinone (5.0 g, 23 mmol) in DMF (11 ml) was added copper cyanide (I) (3.6 g, 39 mmol) and the reaction was heated to 165 ° C. After 6.5 h, the reaction mixture was cooled to room temperature and water (20 ml) and sodium cyanide (3.6 g) were added and the reaction was heated to 100 ° C. After 12 h, the reaction mixture was extracted with ethyl acetate and the combined organic phases were filtered through a short bed of silica gel washing eluting with ethyl acetate. The filtrate was concentrated under reduced pressure to give 1.9 g (51%) of 4-cyano-1,2-benzoxazolinone in the form of a brown solid that did not need further purification. 1 H NMR (400 MHz, DMSO) δ 7.8 (s, 1 H), 7.6 (d, 1 H), 7.2 (d, 1 H).

c) Preparation of N-t-butylacetoxy-4-cyano-1,2-benzoxazolinone

To a solution of 4-cyano-1,2-benzoxazolinone (1.9 g, 15 mmol) in THF (50 ml) at 0 ° C was added triethylamine (2.5 ml, 18 mmol), DMAP (0.37 g, 3.0 mmol) and BOC anhydride (4.3 g, 20 mmol) and the reaction mixture was allowed to warm to room temperature. After 1.5 h, the mixture was quenched with water (100 ml) and extracted with ethyl acetate. Phases combined organics were washed with brine, dried over anhydrous magnesium sulfate and concentrated under reduced pressure giving 4.3 g (100%) of N-t-butylacetoxy-4-cyano-1,2-benzoxazolinone  in the form of a yellow solid that did not need purification additional. 1 H NMR (400 MHz, CDCl 3) δ 7.8 (d, 1 H), 7.55 (d, 1 H), 7.45 (s, 1 H), 1.65 (s, 9 H).

d) Preparation of N-t-butylacetoxy-4-cyano-2-hydroxylaniline

To a solution of N-t-butylacetoxy-4-cyano-1,2-benzoxazolinone (4.3 g, 16 mmol) in methanol (50 ml) potassium carbonate was added (2.3 g, 16 mmol). After 1.5 h, the reaction was stopped with water (100 ml) and extracted with ethyl acetate. Phases combined organics were washed with brine, dried over anhydrous magnesium sulfate and concentrated under reduced pressure giving 3.1 g (81%) of N-t-butylacetoxy-4-cyano-2-hydroxylaniline  in the form of a brown foam that did not need purification additional. 1 H NMR (400 MHz, CDCl 3) δ 7.7 (d, 1 H), 7.15 (s, 1 H), 7.1 (d, 1 H), 1.5 (s, 9 H).

e) Preparation of 4-cyano-2-hydroxylaniline

To a solution of N-t-butylacetoxy-4-cyano-2-hydroxylaniline (3.1 g, 13 mmol) in methylene chloride (100 ml) at 0 ° C was added trifluoroacetic acid and the reaction mixture was allowed to will warm to room temperature. After 2.5 h, the reaction is interrupted with water (100 ml) and extracted with ethyl acetate. The combined organic phases were washed with brine, dried over anhydrous magnesium sulfate and concentrated under pressure reduced The crude material was purified by chromatography of fast column resolution (50% ethyl acetate / hexanes) giving 1.7 g (96%) of 4-cyano-2-hydroxylaniline in the form of a roasted solid. 1 H NMR (400 MHz, DMSO) δ 7.0 (d, 1 H), 6.85 (s, 1 H), 6.65 (d, 1 H); MS (EI) m / e 134 (M +).

f) Preparation of N- (4-cyano-2-hydroxyphenyl) -N '- (2-bromophenyl) thiourea

To a solution of 4-cyano-2-hydroxylaniline (1.0 g, 7.5 mmol) in ethanol (20 ml) was added 2-Bromophenylisothiocyanate (1.0 ml, 7.5 mmol). After 24 h, the reaction mixture was concentrated under pressure. reduced giving 2.0 g (77%) of N- (4-cyano-2-hydroxyphenyl) -N '- (2-bromophenyl) thiourea in the form of a yellow solid that did not need purification additional. 1 H NMR (400 MHz, DMSO) δ 10.8 (s, 1 H), 10.1 (s, 1 H), 9.6 (s, 1 H), 8.7 (d, 1 H), 7.7 (d, 1 H), 7.6 (d, 1 H), 7.4 (t, 1 H), 7.25 (d, 1 H), 7.2 (s, 1 H and d, 2 H); MS (EI) m / e 229 (100), 348 (75 (M +)), 462 (30), 695 (10).

g) Preparation of N- (4-cyano-2-t-butyldimethylsilanoxyphenyl) -N '- (2-bromophenyl) thiourea

To a solution of N- (4-cyano-2-hydroxyphenyl) -N '- (2-bromophenyl) thiourea (3.5 g, 10 mmol) in THF (50 ml) at 0 ° C was added imidazole (1.0 g, 15 mmol) and TBSCl (1.5 g, 10 mmol). After 1 h, the mixture of reaction was quenched with water (100 ml) and extracted with acetate ethyl. The combined organic phases were washed with brine, dried over anhydrous magnesium sulfate and concentrated to reduced pressure The crude material was crystallized from acetate ethyl giving 3.9 g (84%) of N- (4-cyano-2-t-butyldimethylsilanoxyphenyl) -N '- (2-bromophenyl) thiourea  in the form of a yellow solid. 1 H NMR (400 MHz, DMSO) δ 9.9 (s, 1 H), 9.2 (s, 1 H), 8.1 (d, 1 H), 7.7 (d, 1 H), 7.6 (d, 1 H), 7.45 (d, 1 H), 7.4 (t, 1 H), 7.35 (s, 1 H), 7.2 (t, 1 H); MS (EI) m / e 347 (100 (M +)), 175 (40), 461 (40).

h) Preparation of N- (4-cyano-2-t-butyldimethylsilanoxyphenyl) -N '- (2-bromophenyl) carbodiimide

To a solution of N- (4-cyano-2-t-butyldimethylsilanoxyphenyl) -N '- (2-bromophenyl) thiourea (3.4 g, 7.4 mmol) in methylene chloride (40 ml) at 0 ° C was added triethylamine (3.1 ml, 22 mmol), DMAP (20 mg) and chloride methanesulfonyl (1.1 ml, 15 mmol). After 25 min, the mixture of reaction was quenched with water (100 ml) and extracted with acetate ethyl. The combined organic phases were washed with brine, dried over anhydrous magnesium sulfate and concentrated to reduced pressure giving 3.3 g (100%) of N- (4-cyano-2-t-butyldimethylsilanoxyphenyl) -N '- (2-bromophenyl) carbodiimide  in the form of a yellow solid that did not need purification additional. 1 H NMR (400 MHz, DMSO) δ 7.7 (d, 1 H), 7.45 (d, 1 H), 7.4 (m, 4 H), 7.15 (t, 1 H).

i) Conventional procedure for synthesis of cyclic tetraalkylguanidines. Preparation of 4 - [[3- (2-Bromophenyl) -4-oxo-1- (phenylmethyl) -2-imidazolidinyliden] imino] -3-hydroxybenzonitrile

To a solution of N- (4-cyano-2-t-butyldimethylsilanoxyphenyl) -N '- (2-bromophenyl) carbodiimide (86 mg, 0-20 mmol) in THF (2 ml) was added diisopropylethylene (25 µL, 0.57 mmol) and N-benzyl glycine ethyl ester (41 mg, 0.22 mmol). After 15 min, at mixture was added methanol (0.1 ml) and then TBAF (0.24 ml, 0.24 mmol) at 0 ° C. After 30 min, the reaction mixture was quenched with water (2 ml) and extracted with ethyl acetate. Phases combined organics were washed with brine, dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The crude material was purified by recrystallization ( methylene / hexanes) giving 74.5 mg (80%) of 4 - [[3- (2-Bromophenyl) -4-oxo-1- (phenylmethyl) -2-imidazolidinyliden] imino] -3-hydroxybenzonitrile  in the form of a roasted powder. 1 H NMR (400 MHz, MeOD) δ 7.45 (2 H, d), 7.35 (2 H, t), 7.3 (1 H, d), 7.25 (1 H, d), 7.15 (1 H, d), 6.9 (3 H, m), 6.8 (1 H, d), 6.6 (1 H, t), 4.6 (2 H, m), 4.0 (2 H, s); MS (EI) m / e 463 (100 (M +)).

j) Preparation of 4 - [[(7aS) -2- (2-bromophenyl) -hexahydro-1-oxo-3H-pyrrolo [1,2-c] imidazol-3-ylidene] amino] -3-hydroxybenzonitrile

The conventional procedure was followed using N- (4-cyano-2-t-butyldimethylsilanoxyphenyl) -N '- (2-bromophenyl) carbodiimide  (54 mg, 0.13 mmol), diisopropylethylamine (32 µL, 0.29 mmol), L-proline benzyl ester hydrochloride (34 mg, 0.14 mmol) and TBAF (0.16 ml, 0.16 mmol) in THF (2 ml) and methanol (0.1 ml) giving 36 mg (67%) of 4 - [[(7aS) -2- (2-bromophenyl) -hexahydro-1-oxo-3H-pyrrolo [1,2-c] imidazol-3-ylidene] amino] -3-hydroxybenzonitrile  in the form of a roasted powder. 1 H NMR (400 MHz, MeOD) δ 7.6 (2 H, m). 7.35 (3 H, m), 7.15 (1 H, m), 7.0 (1 H, m), 4.5 (1 H, t), 3.0 (1 H, m), 2.4 (1 H, m), 2.2 (1 H, m), 2.05 (2 H, m); MS (EI) m / e 412 (M +).

k) Preparation of 4 - [[5- (2-bromophenyl) -3,3a, 4,5-tetrahydro-1,1-dioxide-4-oxoimidazo [1,5-b] isothiazol-6 (2H) -yliden] amino] - 3-hydroxybenzonitrile

The conventional procedure was followed using N- (4-cyano-2-t-butyldimethylsilanoxyphenyl) -N '- (2-bromophenyl) carbodiimide  (60 mg, 0.14 mmol), diisopropylethylamine (17 µL, 0.15 mmol), aminoester (27 mg, 0.15 mmol) and TBAF (0.17 ml 0.15 mmol) in THF (1.5 ml) and methanol (0.1 ml) giving 46 mg (71%) of 4 - [[5- (2-bromophenyl) -3,3a, 4,5-tetrahydro-1,1-dioxide-4-oxoimidazo [1,5-b] isothiazol-6 (2H) -yliden] amino] - 3-hydroxybenzonitrile  in the form of a roasted powder. 1 H NMR (400 MHz, MeOD) δ 8.1 (1 H, d), 7.8 (1 H, s), 7.75 (1 H, d), 7.7 (1 H, d), 7.6 (2 H, m), 7.25 (1 H, t), 4.35 (1 H, t), 3.9 (2 H, s), 3.25 (1 H, t), 2.9 (1 H, m), 2.7 (1 H, m); MS (EI) m / e 330 (100), 662 (20).

l) Preparation of 4 - [[(6R, 7aS) -2- (2-bromophenyl) -hexahydro-6-hydroxy-1-oxo-3H-pyrrolo [1,2-c] imidazol-3-ylidene] amino] -3-hydroxybenzonitrile

The conventional procedure was followed using N- (4-cyano-2-t-butyldimethylsilanoxyphenyl) -N '- (2-bromophenyl) carbodiimide  (102 mg, 0.24 mmol), diisopropylethylamine (60 µL, 0.26 mmol), L-proline methyl ester hydroxy hydrochloride (44 mg, 0.26 mmol) and TBAF (0.29 ml, 0.29 mmol) in THF (2.5 ml) and methanol (0.1 ml) giving 80 mg (78%) of 4 - [[(6R, 7aS) -2- (2-bromophenyl) -hexahydro-6-hydroxy-1-oxo-3H-pyrrolo [1,2-c] imidazol-3-ylidene] amino] -3-hydroxybenzonitrile in the form of a roasted powder. MS (EI) m / e 427 (100 (M +)).

m) Preparation of 4 - [[(7aR) -2- (2-bromophenyl) -hexahydro-1-oxo-3H-pyrrolo [1,2-c] imidazol-3-ylidene] amino] -3-hydroxybenzonitrile

The conventional procedure was followed using N- (4-cyano-2-t-butyldimethylsilanoxyphenyl) -N '- (2-bromophenyl) carbodiimide  (107 mg, 0.25 mmol), diisopropylethylamine (71 µL, 0.55 mmol), D-proline methyl ester hydrochloride (46 mg, 0.28 mmol) and TBAF (0.30 ml, 0.30 mmol) in THF (2.5 ml) and methanol (0.1 ml) giving 80 mg (78%) of 4 - [[(7aR) -2- (2-bromophenyl) -hexahydro-1-oxo-3H-pyrrolo [1,2-c] imidazol-3-ylidene] amino] -3-hydroxybenzonitrile in the form of a roasted powder. MS (EI) m / e 411 (100 (M +)).

n) Preparation of 4 - [[2- (2-Bromophenyl) -1,5,6,7,8,8a-hexahydro-1-oxoimidazo [1,5-a] pyridin-3 (2H) -iliden] amino] -3- hydroxybenzonitrile

The conventional procedure was followed using N- (4-cyano-2-t-butyldimethylsilanoxyphenyl) -N '- (2-bromophenyl) carbodiimide (106 mg, 0.25 mmol), diisopropylethylamine (71 µL, 0.28 mmol), methylpipecolinate hydrochloride (50 mg, 0.28 mmol) and TBAF (0.30 ml, 0.30 mmol) in THF (2.5 ml) and methanol (0.1 ml) giving 80 mg (75%) from 4 - [[2- (2-Bromophenyl) -1,5,6,7,8,8a-hexahydro-1-oxoimidazo [1,5-a] pyridin-3 (2H) -iliden] amino] -3- hydroxybenzonitrile in the form of a roasted powder. MS (EI) m / e 425 (100 (M +)).

Treatment procedure

The compounds of the invention or a salt pharmaceutically acceptable thereof can be used in the manufacture of a medicament for prophylactic treatment or therapeutic of any pathology in a human being, or another mammal, which worsens or is caused by excessive production or unregulated cytokine IL-8 by the cells of said mammal, such as, but not limited to, monocytes and / or macrophages, or other chemokines that bind to the receptor of IL-8? Or?, Also called type I or type II receiver.

In particular, chemokines are IL-8, GRO?, GRO?, GRO?, NAP-2 or ENA-78.

The compounds of the invention are administered in an amount sufficient to inhibit the function of cytokines, in particular IL-8, GRO?, GRO?, GRO γ, NAP-2 or ENA-78, of so that they are biologically regulated down to levels normal physiological function, or in some cases up to levels below normal, to improve the pathology. The levels abnormal IL-8, GRO?, GRO?, GRO γ, NAP-2 or ENA-78 per example in the context of the present invention, constitute: (i) levels of free IL-8 greater than or equal to 1 picogram per ml; (ii) any cell associated with IL-8, GRO?, GRO?, GRO?, NAP-2 or ENA-78 above normal physiological levels, or (iii) the presence of IL-8, GRO?, GRO?, GRO?, NAP-2 or ENA-78 above basal levels in cells or tissues in IL-8, GRO?, GRO?, GRO?, NAP-2 or ENA-78 respectively.

There are many pathologies in which a production Excessive or unregulated IL-8 is involved in worsen and / or cause disease. Diseases mediated by Chemokines include psoriasis, atopic dermatitis, arthritis, asthma, chronic obstructive pulmonary disease, respiratory failure in adults, inflammatory disease of the intestine, Crohn's disease, ulcerative colitis, accidents cerebrovascular, septic shock, endotoxic shock, sepsis due to gram negatives, toxic shock syndrome, reperfusion injury cardiac and renal, glomerulonephritis, thrombosis, graft reaction against the recipient, Alzheimer's disease, rejections of allografts, malaria, restenosis, angiogenesis, atherosclerosis, osteoporosis, gingivitis and stem cell release unwanted hematopoietic.

These diseases are characterized mainly due to a massive neutrophil infiltration, T cell infiltration, or neovascular growth, and are associated with an increase in the production of IL-8, GRO?, GRO?, GRO? Or NAP-2 which is responsible for neutrophil chemotaxis towards the site inflammatory or directional cell growth endothelial In contrast to other inflammatory cytokines (IL-1, TNF, and IL-6), IL-8, GRO?, GRO?, GRO? Or NAP-2 have the exclusive property of promoting the Neutrophil chemotaxis, release enzymes including, but not limitation, elastase release as well as production and superoxide activation. The α-chemokines, although particularly, GRO?, GRO?, GRO? or NAP-2, which work through the receiver of Type I or II IL-8 can promote neovascularization of tumors promoting directional growth of endothelial cells. Therefore, the inhibition of chemotaxis or activation induced by IL-8 would lead to direct reduction in infiltration of neutrophils

Recent evidence also involves the role of chemokines in the treatment of HIV infections, Littleman et al ., Nature 381, p. 661 (1996) and Koup et al ., Nature 381, p. 667 (1996).

TNF-? Is a cytokine with proinflammatory actions, including the expression of the endothelial leukocyte adhesion molecule. Leukocytes infiltrated in ischemic brain lesions and, therefore, compounds that inhibit or decrease TNF levels will be useful for the treatment of ischemic brain injury. See Liu et al ., Stroke , Vol. 25, No. 7, p. 1481-1488 (1994).

Models of closed cranial lesions and treatment with mixed agents 5-LO / CO are analyzed in Shohami et al ., J. de Vaisc & Clinical Physiology and Pharmacology, Vol. 3, No. 2, p. 99-107 (1992).

The compounds of the invention are administered in an amount sufficient to inhibit binding to IL-8 alpha or beta receptors as evidenced by a reduction in chemotaxis and neutrophil activation. The discovery that the compounds of the invention are inhibitors of IL-8 binding is based on the effects of the compounds of the invention in the in vitro receptor binding assays described herein. The compounds of the invention have proven, in some cases, to be double inhibitors of both recombinant IL-8 type I and type II receptors. Preferably the compounds are inhibitors of only one receptor, more preferably Type II.

As used herein, the expression "IL-8 mediated disease or pathology" is refers to each and every one of the pathologies in which IL-8, GRO?, GRO?, GRO?, NAP-2 or ENA-78 play a role, by production of IL-8 itself, GRO?, GRO?, GRO?, NAP-2 or ENA-78, or because IL-8, GRO?, GRO?, GRO?, NAP-2 or ENA-78 cause the release of another monokine, such as, but not limited to, IL-1, IL-6 or TNF. A pathology in which, for example, IL-1 is a main component, and whose production or action increases or is secreted in response to IL-8, therefore, a pathology mediated by IL-8

As used herein, the expression "chemokine-mediated disease or pathology" refers to each and every one of the pathologies in which a chemokine that is binds to an IL-8? or? receptor plays a role, such as, but not limited to, IL-8, GRO?, GRO?, GRO?, NAP-2 or ENA-78. This would include a pathology in which, IL-8 plays a role, by production of the IL-8 itself, or because IL-8 causes the release of another monokine, such as, but not limited to, IL-1, IL-6 or TNF. A pathology in which, for example, IL-1 is a main component, and whose production or action increases or is secreted in response to IL-8, it would be considered, therefore, a pathology mediated by IL-8

As used in this document, the term "cytokine" refers to any secreted polypeptide that affect the functions of cells and is a molecule that modulates interactions between cells in the immune response, inflammatory or hematopoietic. A cytokine includes, but without limitation, monokines and lymphokines, regardless of what cells produce them. For example, it is generally said that a Monokine is produced and secreted by a mononuclear cell, such as a macrophage and / or a monocyte. However, many others cells also produce monokines, such as killer cells natural, fibroblasts, basophils, neutrophils, cells endothelial, cerebral astrocytes, stromal marrow cells bone, epidermal keratinocytes and B lymphocytes. It is generally said that lymphokines are produced by lymphocytic cells. The Examples of cytokines include, but are not limited to, Interleukin-1 (IL-1), Interleukin-6 (IL-6), Interleukin-8 (IL-8), Factor of Tumor Necrosis-alpha (TNF-?) and Tumor Necrosis Factor-beta (TNF-?).

As used in this document, the term "chemokine" refers to any secreted polypeptide that affect the functions of cells and be a molecule that modulates interactions between cells in the immune response, inflammatory or hematopoietic, similar to the term "cytokine" previous. A chemokine is secreted primarily through cell transmembranes and causes chemotaxis and activation of white blood cells and leukocytes, neutrophils, monocytes, macrophages, T cells, B cells, endothelial cells and muscle cells smooth specific. Examples of chemokines include, but are not limitation, IL-8, GRO-?, GRO-?, GRO-?, NAP-2, ENA-78, IP-10, MIP-1α, MIP-?, PF4, and MCP 1, 2, and 3.

To use a compound of the invention or a salt pharmaceutically acceptable thereof in therapy, usually formulate in a pharmaceutical composition according to practice conventional pharmaceutical This invention, therefore, relates also to a pharmaceutical composition comprising an amount effective, non-toxic, of a compound of the invention and a vehicle or pharmaceutically acceptable diluent.

The compounds of the invention, the salts pharmaceutically acceptable thereof and the compositions pharmaceuticals that incorporate them can be administered conveniently by any of the conventionally used routes  for the administration of drugs, for example, orally, topical, parenteral or by inhalation. The compounds of the invention can be administered in conventional dosage forms prepared by combining a compound of the invention with vehicles conventional pharmacists according to procedures conventional. The compounds of the invention can be administered also in conventional dosages in combination with a second known therapeutically active compound. These procedures may involve mixing, granulation and compression or dissolution of the ingredients as appropriate for the Desired preparation It will be understood that the form and character of the pharmaceutically acceptable carrier or diluent are dictated by the amount of active ingredient with which it is combined, the route of administration and other well known variables. The vehicle or vehicles must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not harmful to the recipient of it.

The pharmaceutical vehicle used can be, by example, a solid or a liquid. Exemplary Solid Vehicles they are lactose, terra alba, sucrose, talc, jelly, agar, pectin, gum arabic, magnesium stearate, stearic acid and the like. Exemplary liquid vehicles are syrup, peanut oil, Olive oil, water Similarly, the vehicle or diluent may include a time-delayed release material well known in the art, such as glyceryl monostearate or Glyceryl distearate alone or with a wax.

A wide variety of ways can be used Pharmaceuticals In this way, if a solid vehicle is attached, the Preparation can be used to form tablets, put in a hard gelatin capsule, in powder or granule form, or in the form of a trocisco or pill. The amount of solid vehicle will vary widely although preferably it will be about 25 mg at approximately 1 g. When a liquid vehicle is used, the Preparation will be in the form of a syrup, emulsion, capsule soft gelatin, a sterile injectable liquid such as a blister or non-aqueous liquid suspension.

The compounds of the invention can administered topically, that is, by administration not systemic This includes the application of a compound of the invention externally to the epidermis or the oral cavity and the instillation of said compound to the ear, eye and nose, so that the compound does not enter the torrent significantly circulatory. By contrast, systemic administration refers to oral, intravenous, intraperitoneal administration and intramuscular

Formulations suitable for administration topical include liquid preparations or semi-liquid suitable for penetration through from the skin to the site of inflammation such as liniments, lotions, creams, ointments or pastes, and drops suitable for administration to eye, ear or nose The active ingredient may comprise, for topical administration, from 0.001% to 10% w / w, for example from 1% to 2% by weight of the Formulation. However, you can understand both as 10% w / w although preferably it will comprise less than 5% w / w, more preferably from 0.1% to 1% w / w of the Formulation.

Lotions in accordance with this invention include those suitable for application to the skin or the eye. An eye lotion may comprise an aqueous solution. sterile that optionally contains a bactericide and can prepare by procedures similar to those for Preparation of drops. Lotions or liniments for application to the skin may also include an agent to accelerate drying and to cool the skin, such as an alcohol or acetone, and / or a humectant such as glycerol or an oil such as castor oil or peanut oil.

Creams, ointments or pastes according to the The present invention are semi-solid formulations of The active ingredients for external application. Can be prepared mixing the active ingredient in a finely divided or powder, alone or in solution or suspension in an aqueous fluid or not aqueous, with the help of suitable machinery, with a greasy base or not grease. The base may comprise hydrocarbons such as paraffin hard, soft or liquid, glycerol, beeswax, a soap metal; a mucilage; an oil of natural origin such as oil of almond, corn, peanut, castor or olive; anhydrous lanolin or its derivatives or a fatty acid such as a steric or oleic acid together with an alcohol such as propylene glycol or a macrogel. The formulation can incorporate any suitable surfactant such as an anionic, cationic or non-ionic surfactant such as a sorbitan ester or a polyoxyethylene derivative thereof. They can also include suspending agents such as natural gums, cellulose derivatives or inorganic materials such as silicas siliceous, and other ingredients such as lanolin.

The drops according to the present invention may comprise aqueous or oily solutions or suspensions sterile and can be prepared by dissolving the active ingredient in a suitable aqueous solution of a bactericidal and / or fungicidal agent and / or any other suitable preservative, and including preferably a surfactant. The resulting solution can then rinse by filtration, transfer to a container suitable that it is then tightly closed and sterilized in autoclave or maintained at 98-100 ° C for half hour. Alternatively, the solution can be sterilized by filtration and transferred to the vessel by a technique aseptic Examples of bactericidal and fungicidal agents suitable for inclusion in the drops are nitrate or acetate phenylmercuric (0.002%), benzalkonium chloride (0.01%) and acetate chlorhexidine (0.01%). Suitable solvents for preparation of an oily solution include glycerol, diluted alcohol and propylene glycol.

The compounds of the invention can administered parenterally, that is, by administration intravenous, intramuscular, subcutaneous, intranasal, intrarectal, intravaginal or intraperitoneal. The subcutaneous forms e intramuscular parenteral administration are preferred usually. The appropriate dosage forms for said Administration can be prepared by conventional techniques. The Compounds of the invention can also be administered by inhalation, that is, by intranasal administration and inhalation oral. The appropriate dosage forms for said administration, such as an aerosol formulation or an inhaler of measured dose, they can be prepared by conventional techniques.

For all the described use procedures in this document for the compounds of the invention the regime of Daily oral dosage will preferably be approximately 0.01 to about 80 mg / kg total body weight. The regime Daily parenteral dosage will be approximately 0.001 a approximately 80 mg / kg total body weight. The regime of daily topical dosage will preferably be from 0.1 mg to 150 mg, administered one to four, preferably two or three times at day. The daily inhalation dosage regimen will be preferably from about 0.01 mg / kg to about 1 mg / kg per day A specialist in the art will also recognize that the optimal amount and the separation of the dosages individual of a compound of the invention or a salt pharmaceutically acceptable thereof will be determined according to the nature and extent of the condition to be treated, the form, route and site of administration, and the particular patient to be treated, and that said Optimal can be determined by conventional techniques. A Technician will also understand that the course optimal treatment, that is, the number of doses of a compound of the invention or a pharmaceutically acceptable salt thereof given per day for a defined number of days, you can set it a specialist in the art using assays to determine the conventional course of treatment.

The invention will now be described by making reference to the following biological examples that are merely illustrative and should not be considered as a scope limitation of the present invention.

Biological examples

The inhibitory effects of IL-8, and the GRO-α chemokine of the compounds of the present invention were determined by the following in vitro assay:

Receiver Binding Assays

[125 I] IL-8 (human recombinant) was obtained from Amersham Corp., Arlington Heights, IL, with specific activity 2000 Ci / mmol. GRO-? Was obtained from NEN-New England Nuclear. The other chemical compounds were of analytical quality. High levels of recombinant human α? And? IL-8 receptors were expressed individually in Chinese hamster ovary cells as described above (Holmes, et al ., Science, 1991, 253, 1278). The Chinese hamster ovary cell membranes were homogenized according to a protocol described above (Haour, et al ., J Biol Chem., 249 p. 2195-2205 (1974)). Except that the homogenization buffer was changed to 10 mM Tris-HCl, 1 mM MgSO 4, 0.5 mM EDTA (ethylenediaminetetraacetic acid), 1 mM PMSF (α-toluenesulfonyl fluoride), 0.5 mg / l of leupeptin, pH 7.5. The membrane protein concentration was determined using a Pierce Co. micro-assay kit using bovine serum albumin as a standard. All tests were performed in a 96-well microplate format. Each reaction mixture contained 125 l IL-8 (0.25 nM) or 125 l GRO-? And 0.5 µg / ml of IL-8R? Or 1.0? Membranes mug / ml of IL-8R? membranes in 20 mM Bis-Trispropane and 0.4 mM Tris HCl buffers, pH 8.0, containing 1.2 mM MgSO4, 0.1 mM EDTA, NaCl 25 mM and 0.03% CHAPS. In addition, drug or compound of interest that had been pre-dissolved in DMSO was added to reach a final concentration of between 0.01 nM and 100 uM. The assay was initiated by the addition of125L-IL-8. After 1 hour at room temperature the plate was collected using a Tomtec 96-well collector on a fiberglass filter blocked with 1% polyethyleneimine / 0.5% BSA and washed 3 times with 25 mM NaCl, TrisHCl 10 mM, 1 mM MgSO 4, 0.5 mM EDTA, 0.03% CHAPS, pH 7.4. The filter was then dried and counted in the Betaplate liquid scintillation counter. The recombinant IL-8Rα or Type I receptor is also referred to herein as the non-permissive receptor and the recombinant IL-8Rβ or Type II receptor is called the permissive receptor.

All the compounds mentioned in this document in the Synthesis Chemistry Section, Example 1 to 15, demonstrated an IC 50 of about 45 to about <1 µg / ml in permissive models for inhibition of IL-8 receptor. Of the compounds tested, he also discovered that Examples 1 to 12 were inhibitors of binding to GRO-? to approximately the same level.

Chemotaxis Assay

The in vitro inhibitory properties of these compounds are determined in the neutrophil chemotaxis assay as described in Current Protocols in Immunology, Vol. I, Supl. 1, Unit 6.12.3. Neutrophils were isolated from human blood as described in Current Protocols in Immunology Vol. I, Supl. 1 Unit 7.23.1. The chemoattractants IL-8, GRO-?, GRO-?, GRO-? And NAP-2 are placed in the lower chamber of a 48 multi-well chamber (Neuro Probe, Cabin John, MD) at a concentration between 0 , 1 and 100 nM. The two chambers are separated by a 5 µm polycarbonate filter. When the compounds of this invention are tested, they are mixed with the cells (0.001-1000 nM) just before the cells are added to the upper chamber. Incubation is allowed to proceed for approximately 45-90 min at approximately 37 ° C in a humidified incubator with 5% CO2. At the end of the incubation period, the polycarbonate membrane is removed and the upper side is washed, then the membrane is stained using the Diff Quick staining protocol (Baxter Products, McGaw Park, IL, USA). Cells that have been chemoattracted by chemokine are counted visually using a microscope. Generally, four fields are counted for each sample, the average of these quantities being calculated to produce the average number of cells that have migrated. Each sample is tested in triplicate and each compound is repeated at least four times. To certain cells (positive control cells) no compound is added, these cells representing the maximum chemotactic response of the cells. In the case where a negative control (not stimulated) is desired, chemokine is not added to the lower chamber. The difference between the positive control and the negative control represents the chemotactic activity of the cells.

Elastase Release Test

The compounds of this invention are tested for their ability to prevent the release of elastase from human neutrophils. Neutrophils are isolated from human blood as described in Current Protocols in Immunology Vol. I, Supl. I Unit 7.23.1. PMN, 0.88 x 10 6 cells, suspended in Ringer's solution (NaCl 118, KCl 4.56, NaHCO 3 25, KH 2 PO 4 1.03, Glucose 11.1 , 5 mM HEPES, pH 7.4) are placed in each well of a 96-well plate in a volume of 50 ul. To this plate is added the test compound (0.001-1000 nM) in a volume of 50 ul, cytochalasin B in a volume of 50 ul (20 ug / ml) and Ringer's buffer in a volume of 50 ul. These cells are allowed to warm (37 ° C, 5% CO2, 95% RH) for 5 min before adding IL-8, GRO?, GRO?, GRO? Or NAP-2 to a final concentration of 0.01 - 1000 nM. The reaction is allowed to proceed for 45 min before centrifuging the 96-well plate (800 xg 5 min) and 100 ul of the supernatant is removed. This supernatant is added to a second 96-well plate followed by an artificial elastase substrate (MeOSuc-Ala-Ala-Pro-Val-AMC, Nova Biochem, La Jolla, CA) to a final concentration of 6 ug / ml dissolved in phosphate buffered saline. Immediately, the plate is placed in a fluorescent 96-well plate reader (Cytofluor 2350, Millipore, Bedfoxd, MA) and the data is collected at 3 min intervals according to the procedure of Nakajima et al. J. Biol. Chem; 254, 4027 (1979). The amount of elastase released from PMNs is calculated by measuring the degradation rate of MeOSuc-Ala-Ala-Pro-Val-AMC.

TNF-? In Brain Injury Assay Traumatic

This essay provides the examination of the mRNA expression of tumor necrosis factor in zones specific brain that appear as a brain injury Traumatic (TBI) with percussion with induced lateral fluid experimentally in rats. Sprague-Dawley Rats adult (n = 42) were anesthetized with sodium pentobarbital (60 mg / kg, i.p.) and underwent a brain injury with fluid percussion lateral of moderate severity (2.4 atm.) centered on the cortex left temporaparietal (n = 18), or "simulated" treatment (anesthesia and surgery without injury, n = 18). The animals are sacrificed by decapitation at 1, 6 and 24 h. After the injury, it they remove the brains, and samples of bark tissue are prepared left parietal (CI) (injured), the corresponding area in the right contralateral cortex (CD), cortex adjacent to the cortex injured parietal (AI), the corresponding adjacent area in the right cortex (AD), left hippocampus (HI) and right hippocampus (HD) Total RNA was isolated and hybridization was performed by Northern blot and was quantified against a control RNA TNF-? positive (macrophage = 100%). Be observes a notable increase in mRNA expression of TNF-? In HI (104 ± 17% of control positive, p <0.05 compared to simulated), CI (105 ± 21%, p <0.05) and AI (69 ± 8%, p <0.01) in the hemisphere with trauma 1 h after injury. There is also an increase in the expression of TNF-? mRNA in HI (46 ± 8%, p <0.05), CI (30 ± 3%, p <0.01) and AI (32 ± 3%, p <0.01) at 6 o'clock which resolves within 24 h following the injury. In the contralateral hemisphere, the TNF-? mRNA expression increases in HD (46 ± 2%, p <0.01), CD (4 ± 3%) and AD (22 ± 8%) in 1 h and in HD (28 ± 11%), CD (7 ± 5%) and AD (26 ± 6%, p <0.05) in 6 h although not in the 24 h after the injury. In animals simulated (surgery without injury) or without prior treatment, it is not observe consistent changes in mRNA expression of TNF-? In any of the 6 brain areas in any of the hemispheres at any time. These results indicate that after a percussion brain injury of parasagittal fluid, the temporal expression of the mRNA of TNF-? Is altered in specific areas of the brain, including those of the hemisphere without trauma. How TNF-? Can induce growth factor nervous (NGF) and stimulate the release of other cytokines to from activated astrocytes, this alteration post-traumatic gene expression of TNF-? Plays an important role in both acute and regenerative responses to CNS trauma.

CNS lesion model for mRNA of IL-?

This essay characterizes regional expression of interleukin-1B mRNA (IL-1B) in specific brain areas after a brain injury Traumatic (TBI) with experimental lateral fluid percussion in rats Adult Sprague-Dawley rats (n = 42) are anesthetize with sodium pentobarbital (60 mg / kg, i.p.) and undergo brain injury with lateral fluid percussion of moderate severity (2.4 atm.) Centered on the left temporaparietal cortex (n = 18), or "simulated" treatment (anesthesia and surgery without injury). The animals are sacrificed at 1, 6 and 24 h after injury, they the brain is removed, and samples of cortex tissue are prepared left parietal (CI) injured, the corresponding area in the right contralateral cortex (CD), cortex adjacent to the cortex injured parietal (AI), the corresponding adjacent area in the right cortex (AD), left hippocampus (HI) and right hippocampus (HD) Total RNA is isolated and hybridization was performed by Northern blot and mRNA amount of IL-1? In brain tissue is presented as percentage with respect to radioactivity of macrophage RNA IL-1B positive that was loaded on the same gel. TO  1 h after brain injury, a noticeable increase is observed and significant in the expression of IL-1B mRNA in CI (20.0 ± 0.7% positive control, n = 6, p <0.05 compared to the simulated animal), HI (24.5 ± 0.9%, p <0.05) and AI (21.5 ± 3.1%, p <0.05) in the injured hemisphere, which remained elevated until 6 h after the IC injury (4.0 ± 0.4%, n = 6, p <0.05) and HI (5.0 ± 1.3%, p <0.05). In simulated animals or without prior treatment, it is not observed IL-1B mRNA expression in any of the areas respective brain. These results indicate that after one TBI, the temporary expression of IL-1B mRNA is It stimulates regionally in specific brain areas. These regional changes in cytokines, such as IL-1 B They play a role after trauma.

Claims (3)

1. A compound selected from the group consisting of: 4 - [[3- (2-Bromophenyl) -4-oxo-1- (phenylmethyl) -2-imidazolidinyliden] imino] -3-hydroxybenzonitrile; 4 - [[(7aS) -2- (2-bromophenyl) -hexahydro-1-oxo-3 H -pyrrolo [1,2-c] imidazol-3-ylidene] amino] -3-hydroxybenzonitrile; 4 - [[5- (2-bromophenyl) -3,3a, 4,5-tetrahydro-1,1-dioxide-4-oxoimidazo [1,5-b] isothiazol-6 (2H) -yliden] amino] - 3-hydro-
xibenzonitrile; 4 - [[(6R, 7aS) -2- (2-bromophenyl) -hexahydro-6-hydroxy-1-oxo-3H-pyrrolo [1,2-c] imidazol-3-ylidene] amino] -3-hydro -
xibenzonitrile; 4 - [[(7aR) -2- (2-bromophenyl) -hexahydro-1-oxo-3H-pyrrolo [1,2-c] imidazol-3-ylidene] amino] -3-hydroxybenzonitrile; 4 - [(S) -2- (2,3-Dichloro-phenyl) -1-oxo-hexahydro-pyrrolo [1,2-c] imidazol-3-ylidenamino] -3-hydroxy-benzonitrile; 4 - [(R) -2- (2,3-Dichloro-phenyl) -1-oxo-hexahydro-pyrrolo [1,2-c] imidazol-3-ylidenamino] -3-hydroxy-benzonitrile; 4- [2- (2,3-Dichloro-phenyl) -1-oxo-hexahydro-pyrrolo [1,2-c] imidazol-3-ylidenamino] -3-hydroxy-benzonitrile; 4 - [(S) -2- (2-Bromo-phenyl) -1-oxo-hexahydro-pyrrolo [1,2-c] imidazol-3-ylidenamino] -3-hydroxy-benzonitrile; 4 - [(R) -2- (2-Bromo-phenyl) -1-oxo-hexahydro-pyrrolo [1,2-c] imidazol-3-ylidenamino] -3-hydroxy-benzonitrile; 4- [2- (2-Bromo-phenyl) -1-oxo-hexahydro-pyrrolo [1,2-c] imidazol-3-ylidenamino] -3-hydroxy-benzonitrile, Y; 4 - [[2- (2-Bromophenyl) -1,5,6,7,8,8a-hexahydro-1-oxoimidazo [1,5-a] pyridin-3 (2H) -iliden] amino] -3- hydroxybenzoni-
trilo; or a pharmaceutically acceptable salt thereof. 2. A pharmaceutical composition that it comprises an effective amount of a compound according to the claim 1, and a pharmaceutical carrier or diluent acceptable. 3. The use of a compound according to claim 1 in the manufacture of a medicament for the treatment of a pathology mediated by selected chemokine between psoriasis, atopic dermatitis, arthritis, asthma, disease chronic obstructive pulmonary, respiratory failure syndrome in adults, inflammatory bowel disease, disease of Crohn, ulcerative colitis, strokes, shock septic, endotoxic shock, gram negative sepsis, syndrome toxic shock, cardiac and renal reperfusion injury, glomerulonephritis, thrombosis, graft reaction against receptor, Alzheimer's disease, allograft rejections, malaria, restenosis, angiogenesis, atherosclerosis, osteoporosis, gingivitis and hematopoietic stem cell release no desired.